Biological Sciences

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    Protocols in Current Issue
    Human Skin Explant Preparation and Culture
    Authors:  Jessica L. Shannon, Stephen J. Kirchner and Jennifer Y. Zhang, date: 09/20/2022, view: 354, Q&A: 0
    [Abstract]

    The ex vivo experimentation with surgically discarded human skin represents a unique methodology amenable for mechanism and pharmacologic agent studies without the involvement of human subjects. Here, we describe a protocol that includes preparation, culture, and stimulation of human skin explants, and subsequent analyses by quantitative reverse

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    Serial Room Temperature Crystal Loading, Data Collection, and Data Processing of Human Glutaminase C in Complex with Allosteric Inhibitors
    Authors:  Shawn K. Milano, D. Marian Szebenyi and Richard A. Cerione, date: 09/20/2022, view: 242, Q&A: 0
    [Abstract]

    Cancer cells often overexpress glutaminase enzymes, in particular glutaminase C (GAC). GAC resides in the mitochondria and catalyzes the hydrolysis of glutamine to glutamate. High levels of GAC have been observed in aggressive cancers and the inhibition of its enzymatic activity has been shown to reduce their growth and survival. Numerous GAC

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    Combination of Sterile Injury and Microbial Contamination to Model Post-surgical Peritoneal Adhesions in Mice
    Authors:  Julia Bayer, Deborah Stroka, Paul Kubes, Daniel Candinas and Joel Zindel, date: 08/20/2022, view: 943, Q&A: 0
    [Abstract]

    Abdominal surgeries are frequently associated with the development of post-surgical adhesions. These are irreversible fibrotic scar bands that appear between abdominal organs and the abdominal wall. Patients suffering from adhesions are at risk of severe complications, such as small bowel obstruction, chronic pelvic pain, or infertility. To date,

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    Aerotaxis Assay in Caenorhabditis elegans to Study Behavioral Plasticity
    Authors:  Qiaochu Li, Daniel-Cosmin Marcu, Paul H. Dear and Karl Emanuel Busch, date: 08/20/2022, view: 608, Q&A: 0
    [Abstract]

    C. elegans shows robust and reproducible behavioral responses to oxygen. Specifically, worms prefer O2 levels of 5–10% and avoid too high or too low O2. Their O2 preference is not fixed but shows plasticity depending on experience, context, or genetic background. We recently showed that this experience-dependent plasticity declines with age,

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    Preparation of a Single-cell Suspension from Drosophila Wing Imaginal Discs
    Authors:  Shu Yang, Brooke Sears and Xiaoyan Zheng, date: 08/20/2022, view: 764, Q&A: 0
    [Abstract]

    The wing imaginal discs in Drosophila larvae are a pair of sac-like structures that later form the wings of the adult fly. During the past decades, wing discs have been used as a simple and accessible model system, for identifying genes and deciphering signaling cascades that play crucial roles in many aspects of development. In this protocol, we

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    Cryptococcus neoformans Virulence Assay Using a Galleria mellonella Larvae Model System
    Authors:  Piotr R. Stempinski, Daniel F. Q. Smith and Arturo Casadevall, date: 08/05/2022, view: 745, Q&A: 0
    [Abstract]

    Cryptococcus neoformans is a human pathogenic fungus that can cause pulmonary infections and meningitis in both immunocompromised and otherwise healthy individuals. Limited treatment options and a high mortality rate underlie the necessity for extensive research of the virulence of C. neoformans. Here we describe a detailed protocol for using the

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    Analysis of Caenorhabditis elegans Aging-related Neurodegeneration in Chemosensory Neurons
    Authors:  Cira Crespo and Roberto Grau, date: 07/20/2022, view: 882, Q&A: 0
    [Abstract]

    Aging and neuronal deterioration constitute important risk factors for the development of neuronal-related diseases, such as different dementia. The nematode Caenorhabditis elegans has emerged as a popular model system for studying neurodegeneration diseases, due to its complete neuronal connectivity map. DiI is a red fluorescent dye that can

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    HaloChIP-seq for Antibody-Independent Mapping of Mouse Transcription Factor Cistromes in vivo
    [Abstract]

    Chromatin immunoprecipitation (ChIP) maps, on a genome-wide scale, transcription factor binding sites, and the distribution of other chromatin-associated proteins and their modifications. As such, it provides valuable insights into mechanisms of gene regulation. However, successful ChIP experiments are dependent on the availability of a

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    Protocol to Study Spatial Subgoal Learning Using Escape Behavior in Mice
    Authors:  Philip Shamash and Tiago Branco, date: 06/20/2022, view: 1092, Q&A: 0
    [Abstract]

    Rodent spatial navigation is a key model system for studying mammalian cognition and its neural mechanisms. Of particular interest is how animals memorize the structure of their environments and compute multi-step routes to a goal. Previous work on multi-step spatial reasoning has generally involved placing rodents at the start of a maze until

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    Time-off-pick Assay to Measure Caenorhabditis elegans Motility
    Authors:  Alyssa C. Walker, Rohan Bhargava, Amanda S. Brust, Ali A. Owji and Daniel M. Czyż, date: 06/20/2022, view: 1345, Q&A: 0
    [Abstract]

    Caenorhabditis elegans is a simple metazoan that is often used as a model organism to study various human ailments with impaired motility phenotypes, including protein conformational diseases. Numerous motility assays that measure neuro-muscular function have been employed using C. elegans. Here, we describe “time-off-pick" (TOP), a novel

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